Paper-based Analytical Devices Research Articles

Colorimetric detection of glucose with smartphone-coupled µPADs: harnessing machine learning algorithms in variable lighting environments

Recent advances in microfluidic paper-based analytical devices (µPADs) have shown immense potential for point-of-care testing (POCT) in resource-constrained settings, especially for monitoring glucose levels in patients with diabetes. However, the practical adoption of smartphone-coupled µPADs has been limited due to the variability in ambient lighting conditions, which affects the colorimetric detection. In this study, we have developed a machine learning-based algorithm together with a flash/no-flash technic, adaptable to various µPADs and capable of accurately monitoring plasma glucose levels across a wide range (25 µM to 30 mM), addressing issues related to environmental illumination variations, and shooting angles.The algorithm is implemented in an Android application, Gluco estimator. It utilizes images captured by smartphones and incorporates intelligent feature engineering. The XYZ color space is adopted for device-independent analysis. KI and TMB color indicators are used to enhance resolution for low and high glucose concentrations, respectively. The EBC classifier, combined with a handcrafted feature set, exhibits outstanding performance, achieving an accuracy of 95% and 91% for TMB and KI, respectively. Additionally, multiple linear regression (MLR) yields high reliability among regression models, with R2 values of 0.95 at 0.25 µM to 3 mM and 0.97 at 3–30 mM. This work represents a significant stride toward realizing a portable, high-resolution, and reliable smartphone-coupled µPAD in uncontrolled environments.

Screening of angiotensin converting enzyme inhibitors from natural products via origami microfluidic paper-based analytical devices with colorimetric detection

We report the screening of angiotensin converting enzyme (ACE) inhibitors on an origami microfluidic paper-based analytical device (μPAD) using colorimetric detection. The hydrolysis product reacts with ninhydrin, resulting in a purple color at the detection zones. Images of the μPADs are captured using a common cell phone and analyzed with Photoshop software. This platform allows six independent colorimetric reactions to take place simultaneously, and the IC50 values can be obtained in a single run within 22 min. The relative standard deviations of inhibition efficiencies are generally lower than 4.0 % (n = 5). The IC50 values of captopril and five products from natural plants were obtained and corresponded well with UV methods. The relative deviations between the two methods are within the range of −5 % to +5 %. This work is a proof-of-concept successfully demonstrating the use of μPADs technology to screen enzyme inhibitors from natural products.

Accelerated naked-eye identification of cations using non-plasmonic amplification by functionalized GQDs: A novel photochemical microfluidic sensor for environmental/mineral analysis using Lab-on-paper technology

This study describes the synthesis of various types of thiolated-graphene quantum dots (GQDs) for the simultaneous multi-sensing of heavy metal ions using one-droplet microfluidic paper-based analytical devices (OD-µPADs). For the first time, functionalized GQDs were synthesized and stabilized on the surface of hydrophobic fiber-glass with simple manufacturing and assembly strategy towards colorimetric chemosensing of ions including iron, copper, cobalt, mercury, chrome, manganese, and nickel. For this purpose, GQDs are functionalized by various organic species including cysteamine, decanethiol, and mercaptopropionic acid (MPA) towards specific electrostatic interaction with metal ions of Fe (II), Cu (II), Co(II), Hg (II), Mn (II), and Ni (II). Therefore, an innovative GQDs-based miniaturized chemo-system was proposed for the environmental analysis. Additionally, the results were confirmed using fluorescence spectrometry as an optical detection method. Finally, this simple and efficient colorimetric platform was used for facile metal ions detection in human urine models, and environmental fluids, which can provide a commercial device for sensing contaminants by non-professional users in biomedical/environmental mixture. As an economical technique, this platform presents a “mix-and-sense” system without dye (as a side substance) or chemical treatments. The low instrumental requirements, the color uniformity, and the disposability of OD-μPADs approve the commercial application of the engineered ion-chemosensor.

Dual Chromatic Laser-Printed Microfluidic Paper-Based Analytical Device (μPAD) for the Detection of Atrazine in Water

Dual Chromatic Laser-Printed Microfluidic Paper-Based Analytical Device (μPAD) for the Detection of Atrazine in Water

Controlling Fluorescent Readout in Paper-based Analytical Devices.

Paper is an ideal candidate for the development of new disposable diagnostic devices because it is a low-cost material, allows transport of the liquid on the device by capillary action, and is environmentally friendly. Today, colorimetric analysis is most often used as a detection method for rapid tests (test strips or lateral flow devices) but usually gives only qualitative results and is limited by a relatively high detection threshold. Here, we describe studies using fluorescence as a readout tool for paper-based diagnostics. We study how the optical readout is affected by light transmission, scattering, and fluorescence as a function of paper characteristics such as thickness (grammage), water content, autofluorescence, and paper type/composition. We show that paper-based fluorescence analysis allows better optical readout compared to that of nitrocellulose, which is currently the material of choice in colorimetric assays. To reduce the loss of analyte molecules (e.g., proteins) due to adsorption to the paper surface, we coat the paper fibers with a protein-repellent hydrogel. For this purpose, we use hydrophilic copolymers consisting of N,N-dimethyl acrylamide and a benzophenone-based cross-linker, which are photochemically transformed into a fiber-attached polymer hydrogel on the paper fiber surfaces in situ. We show that the combination of fluorescence detection and the use of a protein-repellent coating enables sensitive paper-based analysis. Finally, the success of the strategy is demonstrated by using a simple LFD application as an example.

Critical evaluation of analytical methodologies for the discrimination of hemp and drug-type cannabis

Classification of cannabis samples as hemp- or drug-type is an ever-increasing problem worldwide. Colorimetric tests and infrared spectroscopy were selected and critically compared among the available techniques. Cannabis samples with different THC and CBD concentration were analyzed by paper-based analytical devices (µPAD) using the 4-aminophenol colorimetric reaction, attenuated total reflectance mid infrared spectroscopy (ATR-MIR), and diffuse reflectance near infrared spectroscopy (DR-NIR). For the µPAD analysis, cannabis samples were extracted with methanol and later mixed with reagents to develop the colored complex. On the other hand, MIR and NIR spectra were obtained directly on raw samples, and partial least square discriminant analysis (PLS-DA) was used for cannabis classification. All the selected methodologies successfully discriminate between hemp-type and drug-type samples, being suitable to be used in-field. Additionally, assayed methodologies together a reference gas chromatography mass spectrometry (GC–MS) were evaluated and compared from a Green Analytical point of view using the AGREE criteria.

Advancements in Preprocessing and Analysis of Nitrite and Nitrate since 2010 in Biological Samples: A Review.

As a substance present in organisms, nitrite is a metabolite of nitric oxide and can also be ingested. Nitrate is the metabolite of nitrite. Therefore, it is necessary to measure it quickly, easily and accurately to evaluate the health status of humans. Although there have been several reviews on analytical methods for non-biological samples, there have been no reviews focused on both sample preparation and analytical methods for biological samples. First, rapid and accurate nitrite measurement has significant effects on human health. Second, the detection of nitrite in biological samples is problematic due to its very low concentration and matrix interferences. Therefore, the pretreatment plus measuring methods for nitrite and nitrate obtained from biological samples since 2010 are summarized in the present review, and their prospects for the future are proposed. The treatment methods include liquid-liquid microextraction, various derivatization reactions, liquid-liquid extraction, protein precipitation, solid phase extraction, and cloud point extraction. Analytical methods include spectroscopic methods, paper-based analytical devices, ion chromatography, liquid chromatography, gas chromatography-mass spectrometry, electrochemical methods, liquid chromatography-mass spectrometry and capillary electrophoresis. Derivatization reagents with rapid quantitative reactions and advanced extraction methods with high enrichment efficiency are also included. Nitrate and nitrate should be determined at the same time by the same analytical method. In addition, much exploration has been performed on formulating fast testing through microfluidic technology. In this review, the newest developments in nitrite and nitrate processing are a focus in addition to novel techniques employed in such analyses.

Combination of headspace single-drop microextraction (HS-SDME) with a nickel-embedded paper-based analytical device for cyanide quantification

BackgroundCyanide anion can be found in foodstuffs, tobacco smoke and a variety of types of waters, mainly originating from anthropogenic activities. Due to its highly toxic nature, several agencies have established limits for cyanide levels in water. Additionally, monitoring cyanide levels in biological samples, such as blood and urine, is crucial for obtaining clinical information about the health condition of patients. Therefore, there is a pressing need for the development of simple, cost-effective, and reliable analytical methods capable of quantifying cyanide at low concentrations. ResultsThis study presents a novel analytical method for the selective and sensitive determination of cyanide based on analyte volatilization, pre-concentration via single-drop microextraction (SDME) using a selective reagent, and colorimetric quantification using a paper-based analytical device. For this, 10 mL of a liquid sample was acidified with phosphoric acid and the generated HCN was collected using a single drop of 3 μL of a palladium dimethylglyoximate solution (Pd (DMG)22−) positioned in the flask headspace using a syringe. The reaction of Pd (DMG)22− leads to the formation of Pd(CN)42− and the demasking of the organic ligand. After 15 min of extraction time, the reagent drop was added to a paper-based analytical device that has been previously impregnated with 3 μL of nickel chloride, resulting in the formation of a red precipitate of nickel (II) dimethylglyoximate. Digital images of the paper-based device were captured and the red channel (R) was used for quantification purposes. Under optimized conditions, the method demonstrates a suitable linear relation (r2 > 0.99) ranging from 26 to 286 μg L−1 and a limit of detection of 5 μg L−1. SignificanceAs a proof of concept, cyanide levels were quantified in water and urine samples using this method. The proposed approach offers high sensitivity and selectivity while requiring only a small volume of reagents. Furthermore, it exhibits a high degree of portability for in-situ applications.

A sustainable electrochemical analytical platform for rapid detection of indole-3-acetic acid in Marchantia polymorpha

Auxin (mainly indole-3-acetic acid, IAA) is one of the most essential phytohormones with various roles for land plants at different evolution stages and its polar transportation in plants suggested the importance of obtaining its amounts in different plant tissues. Because Marchantia polymorpha is at the entry evolution level, the IAA contents in its tissues would be useful for the study of IAA evolution. Although electrochemical detection offers a possible method for the measurement of IAA amounts, the cost for fabrication of electrochemical sensors is still high given the amounts of IAA tests in the future. Herein a low-cost disposable electrochemical analytical platform was constructed for analysis of IAA in tiny plant samples. Specifically, disposable carbon electrodes were prepared by modifying stainless steel wire mesh with inexpensive carbon ink because of less amounts of materials. The electrode was then integrated with a paper-based analytical device to form a disposable and low-cost platform for rapid electrochemical detection of IAA. For the modified electrode, the highest electrochemical responses for IAA could be obtained when the carbon ink was diluted for 3.5 times. The optimized pH value of the buffer solution for testing was 7.0, which is close to the physiological condition. Coupled with sample treatment using zirconium beads, this approach could successfully differentiate IAA in different tissues of Marchantia polymorpha. The obtained results agreed well with previous reports. This study suggested a sustainable electrochemical analytical detection platform for the study of IAA in practical applications.

A Semi-Automatic Environmental Monitoring Device for Mercury and Cobalt Ion Detection.

A syringe-based, semi-automatic environmental monitoring device is developed for on-site detection of harmful heavy metal ions in water. This portable device consists of a spring-embedded syringe and a polydimethylsiloxane (PDMS) membrane-based flow regulator for semi-automatic fix-and-release fluidic valve actuation, and a paper-based analytical device (PAD) with two kinds of gold nanoclusters (AuNCs) for sensitive Hg2+ and Co2+ ion detection, respectively. The thickness of the elastic PDMS membrane can be adjusted to stabilize and modulate the flow rates generated by the pushing force provided by the spring attached to the plunger. Also, different spring constants can drastically alter the response time. People of all ages can extract the fix-volume sample solutions and then release them to automatically complete the detection process, ensuring high reliability and repeatability. The PAD comprises two layers of modified paper, and each layer is immobilized with bovine serum albumin-capped gold nanoclusters (R-AuNCs) and glutathione-capped gold clusters (G-AuNCs), respectively. The ligands functionalized on the surface of the AuNCs not only can fine-tune the optical properties of the nanoclusters but also enable specific and simultaneous detection of Hg2+ and Co2+ ions via metallophilic Au+ -Hg2+ interaction and the Co2+ -thiol complexation effect, respectively. The feasibility of the device for detecting heavy metal ions at low concentrations in various environmental water samples is demonstrated. The Hg2+ and Co2+ ions can be seen simultaneously within 20 min with detection limits as low as 1.76 nm and 0.27 µm, respectively, lower than those of the regulatory restrictions on water by the US Environmental Protection Agency and the European Union. we expect this sensitive, selective, portable, and easy-to-use device to be valid for on-site multiple heavy metal ion pollution screenings in resource-constrained settings.

Atomically Dispersed Manganese on Graphene Nanosheets as Biocompatible Nanozyme for Glutathione Detection in Liver Tissue Lysate Using Microfluidic Paper-based Analytical Devices.

Recently, single atom catalysts (SACs) featuring M-Nx (M = metal) active sites on carbon support have drawn considerable attention due to their promising enzyme-like catalytic properties. However, typical synthesis methods of SACs often involve energy-intensive carbonization processes. Herein, we report a facile one-pot, low-temperature, wet impregnation method to fully utilize M-N4 sites of manganese phthalocyanine (MnPc) by decorating molecular MnPc over the sheets of graphene nanoplatelets (GNP). The synthesized MnPc@GNP exhibits remarkable peroxidase-mimic catalytic activity toward the oxidation of chromogenic 3,3',5,5'-tetramethylbenzidine (TMB) substrate owing to the efficient utilization of atomically dispersed Mn and the high surface-to-volume ratio of the porous catalyst. A nanozyme-based colorimetric sensing probe is developed to detect important biomarker glutathione (GSH) within only 5 min in solution phase based on the ability of GSH to effectively inhibit the TMB oxidation. The high sensitivity and selectivity of the developed colorimetric assay enable us to quantitatively determine GSH concentration in different biological fluids. This work, for the first time, reports a rapid MnPc@GNP nanozyme-based colorimetric assay in the solid substrate by fabricating microfluidic paper-based analytical devices (μPADs). GSH is successfully detected on the fabricated μPADs coated with only 6.0 μg of nanozyme containing 1.6 nmol of Mn in the linear range of 0.5-10 μM with a limit of detection of 1.23 μM. This work also demonstrates the quantitative detection of GSH in mice liver tissue lysate using μPADs, which paves the way to develop μPADs for point-of-care testing.

Salivary calcium determination with a specially developed microfluidic paper-based device for point-of-care analysis

The calcium monitoring in the body not only anticipates several potential diseases (osteoporosis, kidney stones or high blood pressure) but also helps to improve target therapies and follow-up the patient's health status. Calcium monitoring is essential for the diagnosis of one of the most common endocrine disorders worldwide, namely hyperparathyroidism. So, in this work, a new Point-of-care test (POC-test) using a microfluidic paper-based analytical device (μPAD) for calcium quantification in saliva samples is described. The developed μPAD was based on the colorimetric reaction between calcium and cresolphthalein complexone (CPC) which forms an intense purple colour product. The developed device enabled calcium quantification in the range of 0.27–4.50 mmol/L (11.0–180 mg/L) with a detection limit of 80 µmol/L (3.2 mg/L). The accuracy of the developed μPAD was confirmed by analysing saliva samples (#10) and comparing the results obtained with the atomic absorption spectrometry reference procedure; the relative deviation between the two sets of results was below 10 %. A correlation between salivary calcium content and calcium content in blood was established and it was possible to conclude that salivary calcium concentrations above 1.55 mmol/L is an indicator of hypercalcemia. The developed device was stable for 2 weeks when stored at room temperature in vacuum conditions.

A Double-layered Paper-based Analytical Device for Determination of Iron in Water Samples based on Standard Addition Method

A simple method for the determination of iron involved a novel paper-based analytical device (PAD) was developed. The PAD was composed of two layers. Each layer contained a circular hydrophilic reservoir (10 mm Ø) that was situated in a rectangular filter paper (25 Î 25 mm2). The hydrophobic area was created by painting the paper with a “waterproof” glue. The top and the bottom layers were assigned as “the filtration” and “the detection” platforms, respectively. The procedure was started by pipetting an aliquot of bathophenanthroline (Bphen) onto the hydrophilic zone of the bottom layer followed by spiking of standard solutions (0.1-0.5 mgL-1 Fe2+). The red complex was developed. Then, the top and the bottom layers were assembled by two-sided mounting tape. Later, a water sample was dropped onto the top layer, which removed (filtered) any suspended particles in the water sample. When the filtrate was exposed to the bottom layer, a further colored product formed. The bottom layer was removed and placed in a light-controlled box, and the optical image of the product was captured using a smartphone. Its intensity was evaluated through ImageJTM. Linear standard addition plots were obtained (r2 > 0.99). The PAD provided high precision (RSD < 6%) with good recovery (92.6-102%). It was applied to the analysis of drinking, tap, canal and river water samples without any prior filtration. The iron amounts were compared to the results obtained by the spectrophotometric method, and there was not significantly difference at 95% confidence (Paired-t test, n = 5 samples, tstat = 2.68, tcri = 2.78).

Electroanalytical paper-based device for reliable detection and quantification of sugars in milk

Knowing the level of sugars in food and beverages is essential because they are related to diseases such as diabetes, and obesity, among others. In this regard, an approach has been designed, characterized, and applied to develop the first all-in-one paper-based microfluidic analytical device for the quantification of sugars in milk samples. In particular, the powerful combination of the paper-based platform, the facile manufacturing, and the on-demand customization, allowed to obtain a stand-alone platform that was capable to store all the reagents necessary for the assay, to treating the sample by removing major interferences due to lipids and proteins, and to visualize the results in minutes. A wax-patterned chromatographic paper was finely designed to provide a reagent-free platform for the use of non-specialists applied towards the analysis of sugars' content (lactose as the main sugar). Sugars were electrochemically quantified down to 0.16 g L−1 with a repeatability of 5%. To strengthen the efficacy of the novel platform, the electrochemical paper-based analytical device has been applied to commercial milk, including skimmed, semi-skimmed, whole, and lactose-free, confirming satisfactory performance with analytical reliability. This paper-based architecture opens novel avenues for portable (bio)sensing of other relevant species, not only in the agri-food field.

A Facile Method to Fabricate an Enclosed Paper-Based Analytical Device via Double-Sided Patterning for Ionic Contaminant Detection.

Microfluidic paper-based analytical devices (μPADs) have been developed for use in a variety of diagnosis and analysis fields. However, conventional μPADs with an open-channel system have limitations for application as analytical platforms mainly because of the evaporation and contamination of the sample solution. This study demonstrates the design and fabrication of an enclosed three-dimensional(3D)-μPAD and its application as a primary early analysis platform for ionic contaminants. To generate the hydrophobic PDMS barrier, double-sided patterning is carried out using a PDMS blade-coated stamp mold that is fabricated using 3D printing. The selective PDMS patterning can be achieved with controlled PDMS permeation of the cellulose substrate using 3D-designed stamp molds. We find the optimal conditions enabling the formation of enclosed channels, including round shape pattern and inter-pattern distance of 10 mm of stamp design, contact time of 0.5 min, and spacer height of 300 µm of double-sided patterning procedure. As a proof of concept, this enclosed 3D-μPAD is used for the simultaneous colorimetric detection of heavy metal ions in a concentration range of 0.1-2000 ppm, including nickel (Ni2+), copper (Cu2+), mercury (Hg2+), and radioactive isotope cesium-137 ions (Cs+). We confirm that qualitative analysis and image-based quantitative analysis with high reliability are possible through rapid color changes within 3 min. The limits of detection (LOD) for 0.55 ppm of Ni2+, 5.05 ppm of Cu2+, 0.188 ppm of Hg2+, and 0.016 ppm of Cs+ are observed, respectively. In addition, we confirm that the analysis is highly reliable in a wide range of ion concentrations with CV values below 3% for Ni2+ (0.56%), Cu2+ (0.45%), Hg2+ (1.35%), and Cs+ (2.18%). This method could be a promising technique to develop a 3D-μPAD with various applications as a primary early analysis device in the environmental and biological industries.

Photothermal enhanced pressure sensing based on multifunctional Nb2C MXene for portable detection of interleukin-6

Herein, a sensitive and portable pressure-based biosensor was fabricated to accurately assess maternal plasma interleukin-6 (IL-6) levels for achieving the timely monitoring of preeclampsia. Initially, IL-6 modified Nb2C MXene with excellent physicochemical properties as a multi-functional and powerful biological probe was introduced into the paper-based analytical device via competitive-type immunoassay. As expected, the outstanding peroxidase-like performance of Nb2C MXene triggered the decomposition of hydrogen peroxide and produced the pressure signal. More interestingly, the gas production reaction could be considerably accelerated under the irradiation of 808 nm near-infrared light resulted from the brilliant photothermal characteristic and black-body effect of Nb2C MXene, which induced a significantly enhanced pressure change. Therefore, the synergistic activation and sensitization capabilities of Nb2C MXene on pressure signals make the sensing platform demonstrate satisfactory analytical performances for IL-6 detection including wide linear range from 10−5 to 1 ng/mL, low detection limit of 3.33 × 10−6 ng/mL, high selectivity, good recovery and acceptable repeatability. Evidently, this ingenious strategy not only enriches the application scenarios of MXene, but also provides valuable guidance for developing intelligent and convenient in situ analytical device.

Insights into the Fabrication and Electrochemical Aspects of Paper Microfluidics-Based Biosensor Module.

Over the past ten years, microfluidic paper-based analytical devices (micro-PADs) have attracted a lot of attention as a viable analytical platform. It is expanding as a result of advances in manufacturing processes and device integration. Conventional microfluidics approaches have some drawbacks, including high costs, lengthy evaluation times, complicated fabrication, and the necessity of experienced employees. Hence, it is extremely important to construct a detection system that is quick, affordable, portable, and efficient. Nowadays, micro-PADs are frequently employed, particularly in electrochemical analyses, to replicate the classic standard laboratory experiments on a miniature paper chip. It has benefits like rapid assessment, small sample consumption, quick reaction, accuracy, and multiplex function. The goal of this review is to examine modern paper microfluidics-based electrochemical sensing devices for the detection of macromolecules, small molecules, and cells in a variety of real samples. The design and fabrication of micro-PADs using conventional and the latest techniques have also been discussed in detail. Lastly, the limitations and potential of these analytical platforms are examined in order to shed light on future research.

Flow-Through Carbon Nanofiber-Based Transducer for Inline Electrochemical Detection in Paper-Based Analytical Devices.

Point-of-care (POC) devices are rapid, simple, portable, inexpensive, and convenient, but typically they only deliver qualitative results when used in the form of a lateral flow assay (LFA). Electrochemical detection could improve their sensitivity and ensure quantitative detection; however, a breakthrough in material-based technology is needed. We demonstrate a new concept in which electrodes are directly embedded within the lateral flow, enabling flow-through and hence interaction with the entire sample. This is accomplished through laser-induced carbon nanofibers (LCNFs) made by electrospinning Matrimid into nanofiber mats with subsequent pyrolyzing of electrode structures through a CO2 laser. Their highly porous 3D structure and superior graphene-like electrochemical properties are ideally suited for flow-through electrochemical LFA (EC-LFA), where the LCNFs are simply added in line with the other membranes. After optimization of the setup, biological binding assays typical for LFA diagnostics were successfully implemented, enabling the highly sensitive and quantitative detection of 137 pM DNA target sequences of a pathogenic organism that rivals the performance of pump-controlled microfluidic bioassays. This demonstrates that LCNF-based transducers can transform paper-based diagnostic tests to enable precise, quantitative analysis without reliance on cost-intensive read-out systems.

Maximizing flow rate in single paper layer, rapid flow microfluidic paper-based analytical devices

Small, single-layer microfluidic paper-based analytical devices (µPADs) offer potential for a range of point-of-care applications; however, they have been limited to low flow rates. Here, we investigate the role of laser cutting paper channels in maximizing flow rate in small profile devices with limited fluid volumes. We demonstrate that branching, laser-cut grooves can provide a 59.23–73.98% improvement in flow rate over a single cut, and a 435% increase over paper alone. These design considerations can be applied to more complex microfluidic devices with the aim of increasing the flow rate, and could be used in stand-alone channels for self-pumping.

A paper-based device for glyphosate electrochemical detection in human urine: A case study to demonstrate how the properties of the paper can solve analytical issues

In the ever-growing demand for agricultural production, the use of pesticides and the consequential health risks is an issue that remains in the spotlight. The biomonitoring of pesticides in biological matrices is a mandatory task to point out the adverse effects on those people that are particularly exposed (i.e., occupational exposure) and to customize the use of pesticides for safer and more aware agricultural practices (i.e., precision agriculture). To overcome the bottleneck of costs and long sample treatments, we conceived a paper-based analytical device for the fast and smart detection of glyphosate in human urines, which is still the most widespread pesticide. Importantly, we demonstrate how to face the analytical interference given by uric acid to develop an electrochemical sensor for glyphosate detection using paper as a multifunctional material. To this purpose, a sample treatment was pointed out and integrated into a paper strip to decrease the level of uric acid in urines, finally delivering a ready-to-use device that combines lateral and vertical flow. The effective decrease of uric acid after the paper-integrated treatment is verified by direct oxidation in differential pulse voltammetry, whereas glyphosate detection can be carried out by enzyme inhibition assay in chronoamperometry. The system showed a limit of detection for glyphosate of 75 μg/L and a linear range of 100 - 700 μg/L. Additionally, the sustainability of the paper device was assessed and compared with reference chromatographic methods. Overall, this work provides an example of how to design green sensing solutions for addressing analytical challenges in line with the White Analytical Chemistry principles.